US20190070336A1 - Method for producing extracellular matrix membrane derived from biocompatible porcine cartilage capable of regulating in vivo decomposition rate and physical properties, and composition for preventing adhesion containing extracellular matrix derived from porcine cartilage as active ingredient - Google Patents

Method for producing extracellular matrix membrane derived from biocompatible porcine cartilage capable of regulating in vivo decomposition rate and physical properties, and composition for preventing adhesion containing extracellular matrix derived from porcine cartilage as active ingredient Download PDF

Info

Publication number
US20190070336A1
US20190070336A1 US16/083,564 US201716083564A US2019070336A1 US 20190070336 A1 US20190070336 A1 US 20190070336A1 US 201716083564 A US201716083564 A US 201716083564A US 2019070336 A1 US2019070336 A1 US 2019070336A1
Authority
US
United States
Prior art keywords
extracellular matrix
cartilage
porcine cartilage
porcine
derived
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Abandoned
Application number
US16/083,564
Other languages
English (en)
Inventor
Byoung-Hyun Min
Young Jick KIM
Bo Ram SONG
Hee Woong YUN
Sung In Jeong
Youn-Mook Lim
Jong-Seok Park
Hui-Jeong GWON
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Atems Co Ltd
Original Assignee
Atems Co Ltd
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Atems Co Ltd filed Critical Atems Co Ltd
Assigned to ATEMS CO,. LTD. reassignment ATEMS CO,. LTD. ASSIGNMENT OF ASSIGNORS INTEREST (SEE DOCUMENT FOR DETAILS). Assignors: GWON, Hui-Jeong, JEONG, SUNG IN, KIM, YOUNG JICK, LIM, YOUN-MOOK, MIN, BYOUNG-HYUN, PARK, JONG-SEOK, SONG, Bo Ram, YUN, Hee Woong
Publication of US20190070336A1 publication Critical patent/US20190070336A1/en
Abandoned legal-status Critical Current

Links

Images

Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L31/00—Materials for other surgical articles, e.g. stents, stent-grafts, shunts, surgical drapes, guide wires, materials for adhesion prevention, occluding devices, surgical gloves, tissue fixation devices
    • A61L31/005—Ingredients of undetermined constitution or reaction products thereof
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
    • A61L27/36—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix
    • A61L27/3604—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix characterised by the human or animal origin of the biological material, e.g. hair, fascia, fish scales, silk, shellac, pericardium, pleura, renal tissue, amniotic membrane, parenchymal tissue, fetal tissue, muscle tissue, fat tissue, enamel
    • A61L27/3633—Extracellular matrix [ECM]
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/14—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles
    • A61K9/19—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles lyophilised, i.e. freeze-dried, solutions or dispersions
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L2/00—Methods or apparatus for disinfecting or sterilising materials or objects other than foodstuffs or contact lenses; Accessories therefor
    • A61L2/0005—Methods or apparatus for disinfecting or sterilising materials or objects other than foodstuffs or contact lenses; Accessories therefor for pharmaceuticals, biologicals or living parts
    • A61L2/0011—Methods or apparatus for disinfecting or sterilising materials or objects other than foodstuffs or contact lenses; Accessories therefor for pharmaceuticals, biologicals or living parts using physical methods
    • A61L2/0029—Radiation
    • A61L2/0035—Gamma radiation
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
    • A61L27/36—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
    • A61L27/36—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix
    • A61L27/3604—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix characterised by the human or animal origin of the biological material, e.g. hair, fascia, fish scales, silk, shellac, pericardium, pleura, renal tissue, amniotic membrane, parenchymal tissue, fetal tissue, muscle tissue, fat tissue, enamel
    • A61L27/3612—Cartilage, synovial fluid
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
    • A61L27/36—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix
    • A61L27/3641—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix characterised by the site of application in the body
    • A61L27/3645—Connective tissue
    • A61L27/3654—Cartilage, e.g. meniscus
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
    • A61L27/36—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix
    • A61L27/3683—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix subjected to a specific treatment prior to implantation, e.g. decellularising, demineralising, grinding, cellular disruption/non-collagenous protein removal, anti-calcification, crosslinking, supercritical fluid extraction, enzyme treatment
    • A61L27/3687—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix subjected to a specific treatment prior to implantation, e.g. decellularising, demineralising, grinding, cellular disruption/non-collagenous protein removal, anti-calcification, crosslinking, supercritical fluid extraction, enzyme treatment characterised by the use of chemical agents in the treatment, e.g. specific enzymes, detergents, capping agents, crosslinkers, anticalcification agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
    • A61L27/50—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
    • A61L27/50—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
    • A61L27/52—Hydrogels or hydrocolloids
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
    • A61L27/50—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
    • A61L27/56—Porous materials, e.g. foams or sponges
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L31/00—Materials for other surgical articles, e.g. stents, stent-grafts, shunts, surgical drapes, guide wires, materials for adhesion prevention, occluding devices, surgical gloves, tissue fixation devices
    • A61L31/04—Macromolecular materials
    • A61L31/043—Proteins; Polypeptides; Degradation products thereof
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L31/00—Materials for other surgical articles, e.g. stents, stent-grafts, shunts, surgical drapes, guide wires, materials for adhesion prevention, occluding devices, surgical gloves, tissue fixation devices
    • A61L31/14—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
    • C12N5/06—Animal cells or tissues; Human cells or tissues
    • C12N5/0602—Vertebrate cells
    • C12N5/0652—Cells of skeletal and connective tissues; Mesenchyme
    • C12N5/0655—Chondrocytes; Cartilage
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L2430/00—Materials or treatment for tissue regeneration
    • A61L2430/40—Preparation and treatment of biological tissue for implantation, e.g. decellularisation, cross-linking

Definitions

  • the present invention relates to a method for preparing a biocompatible porcine cartilage-derived extracellular matrix membrane capable of adjusting an in-vovo degradation rate and a mechanical property, and a composition containing a porcine cartilage-derived extracellular matrix as an active ingredient, for preventing adhesion between tissues and/or organs.
  • Natural materials which are derived from natural substances, animals, or human organisms, have very excellent biocompatibility and physiological functions.
  • an extracellular matrix (ECM) may be taken.
  • the extracellular matrix is a biomaterial extracted from complex human and animal tissues, which can control or regulate cell functions or induce tissue regeneration.
  • a supporter made by using a natural material is evaluated as an ideal cell treatment agent or a material for a tissue engineering supporter because it can provide excellent biofunctionality and biodegradability etc., as well as a less inflammatory reaction after transplantation into a living body.
  • techniques have been attracting attention in which allogenic or xenogenic tissues or organs are harvested, and cells are then acellularized and used as various types of supporters or membranes.
  • cartilage-derived extracellular matrix is an anechoic and non-nervous tissue, which is different in tissue specificity from extracellular matrix of other issues.
  • cartilage tissue is rich in chondromodulin, thrombospondin, endostatin, and the like, which inhibit angiogenesis, and further includes lubricin, biglycan, decorin, fibromodulin, etc. which prevent cell adhesion.
  • the extracellular matrix of the cartilage tissue is difficult to adjust a degradation time and its mechanical property is weak, which limits human body application. Accordingly, it is necessary to develop a method of controlling the degradability and improving the mechanical property through physical or chemical treatment thereof.
  • Adhesion is a condition in which tissues of organs to be separated are connected and fused to fibrous tissues by surgical operation or inflammation. Complications thus generated are small bowel obstruction, acquired female infertility, ectopic pregnancy, chronic abdominal pain, reoperation, or the like. The adhesion occurs 50 to 90% after surgical operation, and occurs more often as the number of patients who undergo reoperation by adhesion reaches 34.6%. The adhesion is similar to a healing process that is performed after tissue damage, so it is important to prevent the adhesion without interfering with the healing process.
  • the adhesion occurs when cellulose that is normally deposited during the healing process of a surgical site is continuously developed despite the completion of the healing process, causing it to be combined with adjacent tissues and then ultimately joined to a complete single tissue or organ through penetration of blood vessel. Drugs that accelerate cellulose degradation may be used in order to prevent such adhesion, but in many cases, an anti-adhesion material are used to space tissues by creating physical barriers between the tissues.
  • a biodegradable material such as cellulose or a hyaluronic acid is often used to serve as a physical barrier, or a natural material such as collagen is widely used.
  • a biodegradable material has a property of absorbing water and is characterized by hydrolysis thereof, and is used mainly after abdominal surgery because of their rapid degradation rate.
  • tissue e.g., musculoskeletal tissues, spinal canal, and teeth
  • adhesion inhibitor for each tissue must have necessary properties of the tissue
  • injuries to the musculoskeletal tissues require several weeks or months of regeneration. During this period, adhesion of ligament or tendon are often accompanied. In the case of surgical invasion of joints, rehabilitation exercises over many months are often necessary.
  • the final stage of adhesion is vasculogenesis, and at this stage, an adhered tissue is completely formed into a single tissue, and the adhesion is irreversibly completed. It is therefore very important to prevent formation of blood vessels passing through damaged tissues.
  • the conventional adhesion inhibitors do not have a pharmacological mechanism to prevent ultimate infiltration of blood vessels because they act as simple barriers but have no physiological functions.
  • Patent document 0001 Korean Patent Application Publication No. 10-2010-0041027 (2010.04.22)
  • An aspect of the present invention is to provide a method for preparing a biocompatible porcine cartilage-derived extracellular matrix membrane capable of adjusting an in-vovo degradation rate and a mechanical property, and a composition containing a porcine cartilage-derived extracellular matrix as an active ingredient, for preventing adhesion between tissues and/or organs.
  • An exemplary embodiment of the present invention provides a method for preparing a biocompatible porcine cartilage-derived extracellular matrix membrane capable of adjusting an in-vovo degradation rate and a mechanical property, including: separating porcine cartilage; lyophilizing and pulverizing the separated porcine cartilage; decellularizing the pulverize porcine cartilage powder; preparing an aqueous solution of porcine cartilage powder by mixing the decellularized porcine cartilage powder with an acidic solution and pepsin and then by neutralizing it with a basic solution; preparing a porcine cartilage-derived extracellular matrix membrane by mixing the aqueous solution of porcine cartilage powder with a crosslinking agent; and irradiating the porcine cartilage-derived extracellular matrix membrane with radiation.
  • an exemplary embodiment of the present invention provides a composition containing the porcine cartilage-derived extracellular matrix prepared by the above method as an active ingredient, for preventing adhesion between tissues and/or organs.
  • the present invention relates to a method for preparing a biocompatible porcine cartilage-derived extracellular matrix membrane capable of adjusting an in-vovo degradation rate and a mechanical property, and a composition containing the porcine cartilage-derived extracellular matrix as an active ingredient, for preventing adhesion between tissues and/or organs.
  • cartilage tissue extracellular matrix has difficulty in controlling a decomposition period and its mechanical property is weak, thereby restricting the application. Accordingly, a method of enhancing the mechanical property through physical or chemical treatment and radiation treatment has been developed.
  • biomaterials of various formulations were produced by treating the porcine cartilage-derived extracellular matrix with physiochemical methods.
  • cartilage-specific function was maintained despite the treatment of the physico-chemical treatment was checked.
  • it may also be used as an adhesion inhibitor with excellent in-vivo stability and anti-adhesion effect by using the porcine cartilage-derived extracellular matrix material.
  • FIG. 1 illustrates a preparing process of a porcine cartilage-derived extracellular matrix membrane according to an exemplary embodiment of the present invention.
  • FIG. 2 is a result of comparing before and after crosslinking of the cartilage extracellular matrix membrane.
  • FIG. 3 is a result of analyzing ingredients of a source material of the cartilage extracellular matrix.
  • FIG. 4 is a result of mechanical property analysis by before and after crosslinking of the cartilage extracellular matrix membrane.
  • (a) illustrates a result of tensile strength analysis according to before and after cross-linking and a thickness of the cartilage extracellular matrix membrane.
  • (b) illustrates a result of suture strength measurement according to crosslinking of the cartilage extracellular matrix membrane.
  • FIG. 5 is a result of an enzymatic degradation test according to irradiation of the cartilage extracellular matrix membrane.
  • FIG. 6 is a result of an in-vovo degradation test according to irradiation of the cartilage extracellular matrix membrane.
  • FIG. 7 is a result of a vascular endothelial cell attachment test of the cartilage extracellular matrix membrane.
  • FIG. 8 is a result of an in-vivo effect checking test using a mouse cecum adhesion model.
  • (a) illustrating a mouse cecum adhesion model test process and an extracellular matrix membrane transplantation process.
  • (b) is a comparative photograph with a control group after one week of the extracellular matrix membrane transplantation.
  • (c) illustrates a comparative photograph of histological analysis with a control group after one week of the extracellular matrix membrane transplantation.
  • An exemplary embodiment of the present invention provides a method for preparing a biocompatible porcine cartilage-derived extracellular matrix membrane capable of adjusting an in-vovo degradation rate and a mechanical property, including: separating porcine cartilage; lyophilizing and pulverizing the separated porcine cartilage; decellularizing the pulverize porcine cartilage powder; preparing an aqueous solution of porcine cartilage powder by mixing the decellularized porcine cartilage powder with an acidic solution and pepsin and then by neutralizing it with a basic solution; preparing a porcine cartilage-derived extracellular matrix membrane by mixing the aqueous solution of porcine cartilage powder with a crosslinking agent; and irradiating the porcine cartilage-derived extracellular matrix membrane with radiation.
  • the crosslinking agent is glutaraldehyde, but the prevent invention is not limited thereto.
  • the irradiating is performed by using gamma rays of 5 to 100 KGy, but the prevent invention is not limited thereto.
  • an in-vovo degradation rate and a mechanical property of an in-vivo membrane may be adjusted by treating the porcine cartilage-derived extracellular matrix membrane with radiation, thereby obtaining a sterilization effect.
  • the decellularizing is performed by a physical decellularization method, a chemical decellularization method, or a combination of physical and chemical methods.
  • the physical decellularization method includes freeze-thawing, ultrasonication, or physical stirring.
  • the chemical decellularization method is performed by treating the porcine cartilage-derived powder with a hypotonic buffer, anionic surfactant, non-ionic surfactant, cationic surfactant, DNase, RNase or trypsin.
  • the decellularizing is performed at a temperature range of about 0 to 50° C.
  • the hypotonic buffer may be a Tris HCl (pH 8.0) solution
  • the anionic surfactant may be sodium dodecyl sulfate (SDS), sodium deoxycholate, or Triton X-200
  • the non-ionic surfactant may be Triton X-100
  • the cationic surfactant may be selected from the group consisting of CHAPS, Sulfobetaine-10 (SB-10), Sulfobetaine-16 (SB-16), or Tri-n-butyl phosphate.
  • an exemplary embodiment of the present invention provides a composition containing a porcine cartilage-derived extracellular matrix as an active ingredient, for preventing adhesion, prepared according to the above method.
  • porcine cartilage-derived extracellular matrix may prevent adhesion formation by inhibiting fibrosis and inflammation of the surgical site.
  • composition is in any form selected from the group consisting of ointments, powders, gels, films, slabs, wraps and sponges.
  • porcine knee cartilage of a facility conforming the standard was purchased and used referring to “Animal tissues and their derivatives utilized in the manufacture of medical devices, part 1; Analysis and management of risk, part 2; controls on sourcing, collection and handling” of EN 12442.
  • a preparing process of porcine cartilage powder will be described as follows.
  • a cartilage fragment (about 20 ⁇ 30 mm) was prepared by cutting cartilage from the porcine cartilage, and washed with a physiological saline solution for 10 minutes, frozen at ⁇ 80° C., and lyophilized for 3 days.
  • the lyophilized cartilage fragment was frozen and pulverized to a size of about 10 ⁇ m using a freezing mill (JAI, JFC-300, JAPAN) and stored at ⁇ 80° C.
  • a decellularization process was performed as follows in order to remove cells and genetic materials included in the porcine cartilage powder and obtain pure extracellular matrix components.
  • the porcine cartilage powder prepared was treated with 500 ml of a hypotonic buffer per 10 g and agitated at 200 rpm at 4° C. for 4 hours. In order to precipitate and separate cartilage powder, it was treated at 10,000 rpm for 30 minutes using a centrifugal separator (US-21SMT, Vision, Korea).
  • the decellularized cartilage powder was cooled in a cryocooler of ⁇ 80° C. and lyophilized for 3 days.
  • the dried cartilage powder was pulverized in the same manner as described above to finally obtain the cartilage powder having a size of about 10 ⁇ m and stored at ⁇ 80° C. as needed.
  • Pepsin (Sigma, USA) was treated with 100 ml of a hydrochloric acid aqueous solution per 4 g of decellularized cartilage powder and agitated at 200 rpm at 4° C. for 24 hours.
  • the water-soluble cartilage powder was added to the dialysis membrane (MWCO 1000, Spectrolab, USA) and agitated at 200 rpm at 4° C. for 24 hours in the third distilled water.
  • the water-soluble cartilage powder was placed in a container, cooled in the cryocooler of ⁇ 80° C., and lyophilized for 3 days.
  • the water-soluble cartilage powder was pulverized in the same manner as described above to finally obtain the cartilage powder having a size of about 10 ⁇ m and stored at ⁇ 80° C. as needed.
  • the aqueous solution of cartilage powder was put in a vessel of the centrifugal separator and treated at 3000 rpm for 10 minutes. Using a pipet, the supernatant was dispensed in a volume of 35 ml into a square silicone mold of 100 ⁇ 100 mm and dried in a clean bench for 48 hours.
  • the preparing process of the porcine cartilage extracellular matrix membrane is the same as illustrated in FIG. 1 , and results of the comparison before and after crosslinking of the cartilage extracellular matrix membrane are the same as illustrated in FIG. 2 .
  • the extracellular matrix membrane prepared above was packed with silver foil and irradiated with gamma rays at a dose of 5 KGy to 100 KGy.
  • the cut extracellular matrix membrane was plotted by using a tensile strength meter to show the degree of tensile strength as an ultimate force value. As shown in the results, it is seen that the tensile strength is improved as the thickness of the extracellular matrix increases, and the physical strength is also significantly increased by chemical crosslinking. In addition, it is seen that the physical strength is controllable according to the radiation dose in the measurement of the tensile strength of the sample subjected to the irradiation treatment ( FIG. 4A ).
  • the adhesion-preventing membrane should be fixed to the treatment site, it is effective to prevent the adhesion, so that the membrane is strong enough to be maintained without being broken even when the suture is performed at the time of surgery. Since the suture strength was not specified separately at the
  • the suture strength was measured by cutting the film as follows, sealing the film with a surgical thread, and then measuring the tensile strength by attaching the surgical thread and the extracellular matrix membrane to the tensile strength meter, respectively
  • an adhesion inhibitor having a controlled decomposition period for each organ should be used in order to achieve a specific long-term inhibition effect.
  • Collagenase was treated in vitro and the decomposition behavior according to time was examined in order to examine whether the degree of degradation can be adjusted according to the irradiation doses of the cartilage extracellular matrix membrane. Tests were performed by cutting the cartilage extracellular matrix membrane treated with different doses of radiation by 1 ⁇ 1 cm and then treating it to collagenase-treated PBS to observe it for 2 weeks (Left in FIG. 5 ).
  • the sample treated with collagenase for 2 weeks was centrifuged to analyze the amount of hydroxyproline in the supernatant, such that the degree of degradation of collagen, which is a main ingredient of the extracellular matrix membrane, was measured (Right in FIG. 5 ). As a result of the measurement, it is seen that the degree of degradation of the extracellular matrix membrane was controlled in the treatment of collagenase according to the irradiation dose.
  • Tests were performed as follows in order to check a difference of in-vivo biodegradation according to irradiation of extracellular matrix membrane.
  • a difference of vascular endothelial cell adhesion acting in an adhesion mechanism using the cartilage extracellular matrix membrane prepared in FIG. 1 was checked as follows.
  • Cover glass and the cartilage extracellular matrix membrane prepared in FIG. 1 were attached to a separate 24-well dish having a diameter of 5 mm, dried and fixed, and sterilized by iogas.
  • 2 ⁇ 104 vascular endothelial cells were transplanted onto a film-coated dish, the cover glass, and a non-coated 24-well plate and adhered for 24 hours.
  • the cells attached to each surface were then stained with calein and observed with a fluorescence microscope. As a result, it was seen that cell attachment was inhibited in the dish coated with the extracellular matrix membrane compared to the cover glass and the well plate ( FIG. 7 ).
  • An adhesion model was prepared using a C57BL6 mouse of 8 weeks. After a skin layer and a muscle layer of the abdomen of the mouse were respectively incised and then were sutured to create a model in which adhesion occurs between the damaged tissues. Then, an adhesion effect was checked. Adhesive tissues developed for one week, and the adhesion model in which muscle and/or skin were strongly attached were prepared ( FIG. 8A ).

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Veterinary Medicine (AREA)
  • Epidemiology (AREA)
  • Animal Behavior & Ethology (AREA)
  • Public Health (AREA)
  • Biomedical Technology (AREA)
  • Medicinal Chemistry (AREA)
  • Transplantation (AREA)
  • Dermatology (AREA)
  • Oral & Maxillofacial Surgery (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Botany (AREA)
  • Zoology (AREA)
  • Molecular Biology (AREA)
  • Vascular Medicine (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Surgery (AREA)
  • Heart & Thoracic Surgery (AREA)
  • Dispersion Chemistry (AREA)
  • Genetics & Genomics (AREA)
  • Rheumatology (AREA)
  • Wood Science & Technology (AREA)
  • Biotechnology (AREA)
  • Organic Chemistry (AREA)
  • Urology & Nephrology (AREA)
  • Cell Biology (AREA)
  • General Engineering & Computer Science (AREA)
  • Biophysics (AREA)
  • Biochemistry (AREA)
  • Microbiology (AREA)
  • Pharmacology & Pharmacy (AREA)
  • General Chemical & Material Sciences (AREA)
  • Materials For Medical Uses (AREA)
US16/083,564 2016-03-11 2017-03-09 Method for producing extracellular matrix membrane derived from biocompatible porcine cartilage capable of regulating in vivo decomposition rate and physical properties, and composition for preventing adhesion containing extracellular matrix derived from porcine cartilage as active ingredient Abandoned US20190070336A1 (en)

Applications Claiming Priority (3)

Application Number Priority Date Filing Date Title
KR1020160029579A KR101772316B1 (ko) 2016-03-11 2016-03-11 생체 내 분해속도 및 물성 조절 가능한 생체적합성 돼지연골 유래 세포외기질 막 제조방법 및 상기 돼지연골 유래 세포외기질을 유효성분으로 함유하는 유착방지용 조성물
KR10-2016-0029579 2016-03-11
PCT/KR2017/002563 WO2017155328A1 (ko) 2016-03-11 2017-03-09 생체 내 분해속도 및 물성 조절 가능한 생체적합성 돼지연골 유래 세포외기질 막 제조방법 및 상기 돼지연골 유래 세포외기질을 유효성분으로 함유하는 유착방지용 조성물

Publications (1)

Publication Number Publication Date
US20190070336A1 true US20190070336A1 (en) 2019-03-07

Family

ID=59760222

Family Applications (1)

Application Number Title Priority Date Filing Date
US16/083,564 Abandoned US20190070336A1 (en) 2016-03-11 2017-03-09 Method for producing extracellular matrix membrane derived from biocompatible porcine cartilage capable of regulating in vivo decomposition rate and physical properties, and composition for preventing adhesion containing extracellular matrix derived from porcine cartilage as active ingredient

Country Status (6)

Country Link
US (1) US20190070336A1 (ko)
EP (1) EP3427765A4 (ko)
JP (1) JP6961629B2 (ko)
KR (1) KR101772316B1 (ko)
CN (1) CN108834404A (ko)
WO (1) WO2017155328A1 (ko)

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US10939362B2 (en) 2018-01-11 2021-03-02 Mediatek Inc. Apparatuses and methods for system information (SI) request through a contention-based random access procedure
WO2022093877A1 (en) * 2020-10-26 2022-05-05 Briopryme Biologics, Inc. Preparation and use of tissue matrix derived powder

Families Citing this family (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR102038434B1 (ko) * 2018-01-16 2019-10-30 한국원자력연구원 동물의 연골 유래 세포외기질 막의 분해도 조절 방법 및 이를 이용하여 분해도가 조절된 동물의 연골 유래 세포외기질 막
KR102333417B1 (ko) 2019-08-27 2021-12-02 주식회사 도프 동물지방 유래 세포외기질 및 동물지방 유래 세포외기질 보존액
KR102416861B1 (ko) * 2020-02-28 2022-07-07 주식회사 엘앤씨바이오 연골 성분을 함유하는 연골 재생용 조성물 및 그 제조방법
US20230121257A1 (en) * 2020-02-28 2023-04-20 L&C Bio Co., Ltd. Composition comprising cartilage ingredient for regeneration of cartilage and preparation method therefor
CN111840642B (zh) * 2020-07-15 2022-04-01 四川大学 一种软骨脱细胞基质复合支架的制备方法及其应用
KR102549899B1 (ko) * 2021-02-19 2023-07-03 한국원자력연구원 동물의 연골 유래 세포외기질 막의 접착력 조절 방법 및 이를 이용하여 접착력이 조절된 동물의 연골 유래 세포외기질 막

Family Cites Families (14)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4633558A (en) * 1986-02-27 1987-01-06 Wittek Industries, Inc. Cam action tool for applying a spring clamp
JPH01502081A (ja) * 1987-01-15 1989-07-27 レスカ−デン　インコ−ポレ−テツド 動物軟骨の加工方法
JP3536295B2 (ja) * 1999-01-21 2004-06-07 ニプロ株式会社 縫合可能な組織再生誘導型癒着防止膜
JP4595279B2 (ja) * 2001-12-13 2010-12-08 ニプロ株式会社 癒着防止膜
KR101056069B1 (ko) * 2008-10-13 2011-08-10 아주대학교산학협력단 동물조직 분말을 이용한 다공성 3차원 지지체의 제조방법
JP2010279574A (ja) * 2009-06-05 2010-12-16 Nipro Corp 癒着防止膜
KR101269618B1 (ko) * 2011-04-12 2013-06-05 한스바이오메드 주식회사 포유류의 연골조직에서 유래한 생체이식재
CN102188748B (zh) * 2011-04-29 2014-02-26 成都军区昆明总医院 一种新型软骨细胞外基质膜及其制备方法
KR20130134447A (ko) * 2012-05-31 2013-12-10 리젠프라임 주식회사 신규한 나노섬유막 및 이의 제조방법
GB201215725D0 (en) * 2012-09-04 2012-10-17 Univ Leeds Composite connective tissue and bone implants
US10722614B2 (en) * 2013-09-25 2020-07-28 The University Of Kansas Decellularized hyaline cartilage powder for tissue scaffolds
KR20150067518A (ko) * 2013-12-10 2015-06-18 인제대학교 산학협력단 연골세포 유래 세포외 기질막을 유효성분으로 함유하여 수술 후 유착 방지용 조성물
US10213526B2 (en) * 2014-03-21 2019-02-26 University Of Pittsburgh-Of The Commonwealth System Of Higher Education Methods for preparation of a terminally sterilized hydrogel derived from extracellular matrix
KR101633585B1 (ko) * 2014-03-31 2016-06-27 아주대학교산학협력단 표면개질된 연골세포 유래 세포외 기질막을 유효성분으로 함유하는 유착방지용 조성물

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US10939362B2 (en) 2018-01-11 2021-03-02 Mediatek Inc. Apparatuses and methods for system information (SI) request through a contention-based random access procedure
WO2022093877A1 (en) * 2020-10-26 2022-05-05 Briopryme Biologics, Inc. Preparation and use of tissue matrix derived powder
EP4232050A4 (en) * 2020-10-26 2024-05-08 Biopryme Biologics, Inc. PREPARATION AND USE OF POWDER DERIVED FROM TISSUE MATRIX

Also Published As

Publication number Publication date
CN108834404A (zh) 2018-11-16
JP2019508213A (ja) 2019-03-28
KR101772316B1 (ko) 2017-08-29
WO2017155328A1 (ko) 2017-09-14
JP6961629B2 (ja) 2021-11-05
EP3427765A4 (en) 2020-01-15
EP3427765A1 (en) 2019-01-16

Similar Documents

Publication Publication Date Title
US20190070336A1 (en) Method for producing extracellular matrix membrane derived from biocompatible porcine cartilage capable of regulating in vivo decomposition rate and physical properties, and composition for preventing adhesion containing extracellular matrix derived from porcine cartilage as active ingredient
US11083823B2 (en) Tissue-separating fatty acid adhesion barrier
DK2121048T3 (en) Hemostatic compositions and therapeutic regimens
KR20170003916A (ko) 세포외 기질로부터 유래한 최종 멸균된 하이드로겔의 제조 방법
JP2010533042A (ja) 組織修復用無細胞組織基質構成体
CN106163531B (zh) 将表面改性的软骨细胞衍生细胞外基质膜作为有效成分包含的组合物在防止粘连中的应用
US20210393396A1 (en) Dermal layer for grafting having improved graft survival rate and method for producing same
US20220339324A1 (en) Acellular artificial skin substitute and method of preparation thereof
Chou et al. Bioabsorbable fish scale for the internal fixation of fracture: a preliminary study
Zhang et al. Additively manufactured macroporous chambers facilitate large volume soft tissue regeneration from adipose-derived extracellular matrix
JP6382854B2 (ja) ＩｎＳｉｔｕで重合する注射可能なコラーゲン組成物
KR20170110882A (ko) 장기 유착 방지용 수화겔 및 이를 이용한 수화겔 필름
US20100015226A1 (en) Bioresorbable hydrogel
AU2015397501B2 (en) Method for manufacturing collagen film using ultraviolet light, collagen film manufactured by using same, and biomaterial prepared using collagen film
JP2009513290A (ja) 強膜バックリングバンドとその製造方法
Kumar et al. Acellular dermal grafts for the reconstruction of umbilical/ventral hernias in buffalo calves
KR20180029021A (ko) 장기 유착 방지용 수화겔 및 이를 이용한 수화겔 필름
RU2766191C1 (ru) Способ применения биополимерной пленки на основе Acetobacter xylinum для протезирования мягких тканей животных
He et al. Integrity of the ECM influences the bone regenerative property of ECM/dicalcium phosphate composite scaffolds
AU2015264846B2 (en) Hemostatic compositions and therapeutic regimens
Xu et al. Ox Horn biocompatibility and biosafety are sufficient for use in clinical orthopedics
CN109847111A (zh) 一种包含白及多糖的防粘连材料及其制备方法
Vineet Kumar et al. Acellular dermal grafts for the reconstruction of umbilical/ventral hernias in buffalo calves.

Legal Events

Date Code Title Description
AS Assignment

Owner name: ATEMS CO,. LTD., KOREA, REPUBLIC OF

Free format text: ASSIGNMENT OF ASSIGNORS INTEREST;ASSIGNORS:MIN, BYOUNG-HYUN;KIM, YOUNG JICK;SONG, BO RAM;AND OTHERS;REEL/FRAME:046824/0817

Effective date: 20180910

STPP Information on status: patent application and granting procedure in general

Free format text: APPLICATION DISPATCHED FROM PREEXAM, NOT YET DOCKETED

STPP Information on status: patent application and granting procedure in general

Free format text: DOCKETED NEW CASE - READY FOR EXAMINATION

STPP Information on status: patent application and granting procedure in general

Free format text: NON FINAL ACTION MAILED

STPP Information on status: patent application and granting procedure in general

Free format text: RESPONSE TO NON-FINAL OFFICE ACTION ENTERED AND FORWARDED TO EXAMINER

STPP Information on status: patent application and granting procedure in general

Free format text: FINAL REJECTION MAILED

STPP Information on status: patent application and granting procedure in general

Free format text: ADVISORY ACTION MAILED

STCB Information on status: application discontinuation

Free format text: ABANDONED -- FAILURE TO RESPOND TO AN OFFICE ACTION